摘要
对含伪狂犬病病毒（Pseudorabies Virus,PrV）Ea株gD基因部分编码序列,gI、gE和11k基因全序列、28k基因部分序列的质粒pSKB4.5进行亚克隆,构建了只含完整gE基因（长1.78kb）的重组质粒pSDM1.78+,并采用双脱氧末端终止法对全序列进行了分析,发现同国外标准毒株Rice株相比较,在核苷酸和氨基酸水平均存在一定程度的差异。进一步将gE基因克隆到高效真核表达载体pcDNA3.1+的KpnI和BamHI位点之间,构建了gE基因的真核表达质粒pcDNA-gE。体外转染 IBRS-2细胞,经间接免疫荧光法检测证实了gE基因在IBRS-2细胞中得到了表达,表达的蛋白具有生物学活性。
A recombinant plasmid pSDM1. 78 + containing the complete glycoprotein E gene of pseudorabies virus (PRV) Ea strain was constructed by subcloning from plasmid pSKB4.5, which was composed of the gI, gE,11K genes and partial sequence of gD,28K genes. The results of sequence analysis showed that there were multiple mutants compared with PRV Rice strain. The gE gene was further cloned into the KpnI and BamHI sites of eukaryotic expression vector pcDNA3.1 + ,resuting in a eukaryotic expression plasmid pcDNA-gE. This expression plasmid was transfected into the IBRS-2 cell and the cell lines expressed stablely glycoprotein E was established. Expression protein distributed on the membrance and had biology activity by Indirect Immunoflures-ence Assay.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2002年第2期174-179,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
"九五"国家科技攻关计划生物技术项目(96-C01-04-03)
